Photonic crystal fiber (PCF) designs with two zero-dispersion wavelengths (ZDWs) are experimentally investigated in order to suggest a novel PCF for coherent anti-Stokes Raman scattering (CARS) microscopy. From our investigation, we select the optimum PCF design and demonstrate a tailored spectrum with power concentrated around the relevant wavelengths for lipid imaging (648 nm and 1027 nm). This new PCF is characterized by varying the fiber length, the average power, and the pulse width of the fs pump pulses. It was found that the selected PCF design gave a significantly improved spectral distribution compared to an existing PCF for CARS microscopy. Furthermore, the PCF is designed in a twofold symmetric structure allowing for polarization maintaining propagation. Finally, the pulse propagation is investigated numerically showing good agreement with the measured spectrum. From the numerical analysis, the nonlinear effects responsible for the spectral broadening are explained to be soliton fission processes, dispersive waves, and stimulated Raman scattering.